*Target (protein/gene name): Thioredoxin reductase *NCBI Gene # or RefSeq#: TDR Targets ID: 9997, GeneID: 886232 *Protein ID (NP or XP #) or Wolbachia#:83753940/83753941 *Organism (including strain): Mycobacterium tuberculosis Etiologic Risk Group (see link below): Risk Group 3 (RG3) - Bacterial Agents Including Rickettsia *Background/Disease Information (sort of like the Intro to your Mini Research Write up):
Essentiality of this protein: essential Complex of proteins?: no Druggable Target: 0.5 on druggability index
*EC#: 1.6.4.5/1.8.1.9 (TDR Targets lists both) Link to BRENDA EC# page: 1.8.1.9 (transferred from 1.6.4.5) --Show screenshot of BRENDA enzyme mechanism schematic Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): -- link to Sigma (or other company) page for assay or assay reagents (substrates) -- link (or citation) to paper that contains assay information -- List cost and quantity of substrate reagents and supplier
-Potassium phosphate monobasic, 100g ($19.90 from Sigma-Aldrich)
-Ethylenediaminetetraacetic acid disodium salt dihydrate, 50g ($24.50 from Sigma)
-β-Nicotinamide adenine dinucleotide 2′-phosphate reduced tetrasodium salt hydrate, 25mg ($85.10 from Sigma)
-Albumin from bovine serum, 10g ($73.60 from Sigma)
-Thioredoxin, 1mg ($173.00 from Sigma)
-100% Ethanol 500ml ($71.00 from Sigma)
-5,5′-Dithiobis(2-nitrobenzoic acid), 500mg ($15.70 from Sigma)
Structure Available (PDB or Homology model)
-- PDB # or closest PDB entry if using homology model: 2A87
-- For Homology Model option:
---- Show pairwise alignment of your BLASTP search in NCBI against the PDB
---- Query Coverage:
---- Max % Identities:
---- % Positives
---- Chain used for homology:
Current Inhibitors: Expression Information (has it been expressed in bacterial cells):expressed in E. coli Purification Method:Using C-terminus 6His tag Image of protein (PyMol with features delineated and shown separately): *Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
*length of your protein in Amino Acids 335 Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website: 35643.0 kDa Molar Extinction coefficient of your protein at 280 nm wavelength: 14565. assuming all pairs of Cys residues form cystines TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
*CDS Gene Sequence (paste as text only):
ATGACCGCCCCGCCTGTCCATGACCGCGCACACCACCCCGTTCGCGACGTGATCGTTATCGGCTCCGGTCCCGCGGGGTACACTGCGGCGCTCTACGCCGCCCGTGCCCAGCTGGCGCCGCTGGTCTTCGAGGGCACGTCTTTCGGCGGCGCGCTGATGACCACCACCGACGTGGAGAACTACCCGGGATTTCGCAACGGCATCACCGGTCCAGAGTTGATGGATGAGATGCGGGAACAGGCGCTGCGATTCGGCGCGGACCTGCGTATGGAAGACGTCGAGTCGGTATCACTTCACGGGCCGCTGAAATCGGTCGTCACCGCCGACGGACAGACCCACCGGGCCCGAGCCGTGATCCTGGCAATGGGCGCAGCGGCACGCTATCTGCAGGTGCCCGGCGAACAGGAATTGCTCGGGCGCGGGGTGAGCTCGTGCGCCACCTGCGACGGATTCTTCTTCCGCGATCAGGACATCGCCGTCATCGGCGGCGGTGACTCGGCAATGGAGGAAGCTACCTTCCTGACCCGATTCGCTCGCAGTGTGACGCTGGTGCATCGCCGCGACGAGTTCCGGGCTTCCAAAATCATGCTCGATCGCGCCCGCAACAACGACAAGATACGGTTCCTCACCAACCACACCGTGGTCGCGGTGGACGGGGACACCACAGTGACCGGCTTGCGGGTACGCGACACCAACACCGGTGCCGAAACCACCCTGCCGGTAACCGGTGTTTTCGTCGCGATCGGCCACGAGCCGCGGTCGGGCTTGGTGCGCGAGGCCATCGACGTCGACCCGGACGGCTACGTGTTGGTGCAGGGGCGTACCACCAGCACCTCACTGCCGGGCGTGTTCGCTGCCGGCGACCTGGTGGATCGCACCTATCGCCAGGCGGTTACCGCAGCGGGCAGTGGCTGCGCCGCGGCTATCGACGCCGAGCGCTGGCTCGCCGAGCACGCAGCAACCGGAGAAGCTGACAGTACCGACGCATTGATAGGAGCACAACGATGA *GC% Content for gene: 66.1% *CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only): *GC% Content for gene (codon optimized):
Do Not Need this info for Spring (but still copy these lines to your Target page for now) Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): (link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences): TDR Targets ID: 9997
*NCBI Gene # or RefSeq#: TDR Targets ID: 9997, GeneID: 886232
*Protein ID (NP or XP #) or Wolbachia#: 83753940/83753941
*Organism (including strain): Mycobacterium tuberculosis
Etiologic Risk Group (see link below): Risk Group 3 (RG3) - Bacterial Agents Including Rickettsia
*Background/Disease Information (sort of like the Intro to your Mini Research Write up):
Essentiality of this protein: essential
Complex of proteins?: no
Druggable Target: 0.5 on druggability index
*EC#: 1.6.4.5/1.8.1.9 (TDR Targets lists both)
Link to BRENDA EC# page: 1.8.1.9 (transferred from 1.6.4.5)
-- Show screenshot of BRENDA enzyme mechanism schematic
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):
-- link to Sigma (or other company) page for assay or assay reagents (substrates)
-- link (or citation) to paper that contains assay information
-- List cost and quantity of substrate reagents and supplier
-Potassium phosphate monobasic, 100g ($19.90 from Sigma-Aldrich)
-Ethylenediaminetetraacetic acid disodium salt dihydrate, 50g ($24.50 from Sigma)
-β-Nicotinamide adenine dinucleotide 2′-phosphate reduced tetrasodium salt hydrate, 25mg ($85.10 from Sigma)
-Albumin from bovine serum, 10g ($73.60 from Sigma)
-Thioredoxin, 1mg ($173.00 from Sigma)
-100% Ethanol 500ml ($71.00 from Sigma)
-5,5′-Dithiobis(2-nitrobenzoic acid), 500mg ($15.70 from Sigma)
Structure Available (PDB or Homology model)
-- PDB # or closest PDB entry if using homology model: 2A87
-- For Homology Model option:
---- Show pairwise alignment of your BLASTP search in NCBI against the PDB
---- Query Coverage:
---- Max % Identities:
---- % Positives
---- Chain used for homology:
Current Inhibitors:
Expression Information (has it been expressed in bacterial cells): expressed in E. coli
Purification Method: Using C-terminus 6His tag
Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
1 mtappvhdra hhpvrdvivi gsgpagytaa lyaaraqlap lvfegtsfgg almtttdven
61 ypgfrngitg pelmdemreq alrfgadlrm edvesvslhg plksvvtadg qthraravil
121 amgaaarylq vpgeqellgr gvsscatcdg fffrdqdiav igggdsamee atfltrfars
181 vtlvhrrdef raskimldra rnndkirflt nhtvvavdgd ttvtglrvrd tntgaettlp
241 vtgvfvaigh eprsglvrea idvdpdgyvl vqgrttstsl pgvfaagdlv drtyrqavta
301 agsgcaaaid aerwlaehaa tgeadstdal igaqr
*length of your protein in Amino Acids 335
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website: 35643.0 kDa
Molar Extinction coefficient of your protein at 280 nm wavelength: 14565. assuming all pairs of Cys residues form cystines
TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
*CDS Gene Sequence (paste as text only):
ATGACCGCCCCGCCTGTCCATGACCGCGCACACCACCCCGTTCGCGACGTGATCGTTATCGGCTCCGGTCCCGCGGGGTACACTGCGGCGCTCTACGCCGCCCGTGCCCAGCTGGCGCCGCTGGTCTTCGAGGGCACGTCTTTCGGCGGCGCGCTGATGACCACCACCGACGTGGAGAACTACCCGGGATTTCGCAACGGCATCACCGGTCCAGAGTTGATGGATGAGATGCGGGAACAGGCGCTGCGATTCGGCGCGGACCTGCGTATGGAAGACGTCGAGTCGGTATCACTTCACGGGCCGCTGAAATCGGTCGTCACCGCCGACGGACAGACCCACCGGGCCCGAGCCGTGATCCTGGCAATGGGCGCAGCGGCACGCTATCTGCAGGTGCCCGGCGAACAGGAATTGCTCGGGCGCGGGGTGAGCTCGTGCGCCACCTGCGACGGATTCTTCTTCCGCGATCAGGACATCGCCGTCATCGGCGGCGGTGACTCGGCAATGGAGGAAGCTACCTTCCTGACCCGATTCGCTCGCAGTGTGACGCTGGTGCATCGCCGCGACGAGTTCCGGGCTTCCAAAATCATGCTCGATCGCGCCCGCAACAACGACAAGATACGGTTCCTCACCAACCACACCGTGGTCGCGGTGGACGGGGACACCACAGTGACCGGCTTGCGGGTACGCGACACCAACACCGGTGCCGAAACCACCCTGCCGGTAACCGGTGTTTTCGTCGCGATCGGCCACGAGCCGCGGTCGGGCTTGGTGCGCGAGGCCATCGACGTCGACCCGGACGGCTACGTGTTGGTGCAGGGGCGTACCACCAGCACCTCACTGCCGGGCGTGTTCGCTGCCGGCGACCTGGTGGATCGCACCTATCGCCAGGCGGTTACCGCAGCGGGCAGTGGCTGCGCCGCGGCTATCGACGCCGAGCGCTGGCTCGCCGAGCACGCAGCAACCGGAGAAGCTGACAGTACCGACGCATTGATAGGAGCACAACGATGA
*GC% Content for gene: 66.1%
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
*GC% Content for gene (codon optimized):
Do Not Need this info for Spring (but still copy these lines to your Target page for now)
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences):
TDR Targets ID: 9997